GCK-MODY diabetes associated with protein misfolding, cellular self-association and degradation

Department of Clinical Medicine, University of Bergen, N-5020 Bergen, Norway.
Biochimica et Biophysica Acta (Impact Factor: 4.66). 07/2012; 1822(11):1705-15. DOI: 10.1016/j.bbadis.2012.07.005
Source: PubMed


GCK-MODY, dominantly inherited mild fasting hyperglycemia, has been associated with >600 different mutations in the glucokinase (GK)-encoding gene (GCK). When expressed as recombinant pancreatic proteins, some mutations result in enzymes with normal/near-normal catalytic properties. The molecular mechanism(s) of GCK-MODY due to these mutations has remained elusive. Here, we aimed to explore the molecular mechanisms for two such catalytically 'normal' GCK mutations (S263P and G264S) in the F260-L270 loop of GK. When stably overexpressed in HEK293 cells and MIN6 β-cells, the S263P- and G264S-encoded mutations generated misfolded proteins with an increased rate of degradation (S263P>G264S) by the protein quality control machinery, and a propensity to self-associate (G264S>S263P) and form dimers (SDS resistant) and aggregates (partly Triton X-100 insoluble), as determined by pulse-chase experiments and subcellular fractionation. Thus, the GCK-MODY mutations S263P and G264S lead to protein misfolding causing destabilization, cellular dimerization/aggregation and enhanced rate of degradation. In silico predicted conformational changes of the F260-L270 loop structure are considered to mediate the dimerization of both mutant proteins by a domain swapping mechanism. Thus, similar properties may represent the molecular mechanisms for additional unexplained GCK-MODY mutations, and may also contribute to the disease mechanism in other previously characterized GCK-MODY inactivating mutations.

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Available from: Torgeir Flatmark, Jul 20, 2014
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    • "The structural effects of the mutation are related to those previously observed for two mutations in the loop structure itself (S263P and G264S) which, contrary to the R275C mutant, demonstrated a reduced catalytic efficiency. The molecular mechanism proposed for the formation of a high proportion of dimers, when S263P and G264S are expressed in cells (Negahdar et al., 2012), also apply for the R275C mutant protein. "
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